JP2001354692A - Method for producing cytidine derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an effective method for producing a cytidine derivative. SOLUTION: This method for producing the cytidine derivative expressed by formula (3) comprises reacting a uridine derivative expressed by formula (1) with a tertiary amine together with a reagent having a dehydrating property and then with ammonia, or a primary or secondary amine expressed by formula (2). (In formulae (1), (2), (3), X is H, a halogen atom, a 1-4C alkyl or the like; R1, R2 are each independently H, or a blocking group for hydroxyl group; R3 is H, a halogen atom, hydroxyl, a 1-4C alkyl, cyano, an alkenyl, an alkynyl, a 1-4C alkoxyl or the like; R4, R5 are each independently H, a 1-4C alkyl, a 5-8C cycloalkyl, a 1-4C alkyl group substituted by a halogen atom or a 2-4C alkenyl; and R4 and R5 may form a ring by binding them together).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing a cytidine derivative. A cytidine derivative is a compound that is useful as a pharmaceutical agent and an agricultural chemical including an anticancer agent and an antiviral agent, and is also useful as a raw material for antisense DNA and the like that has been recently developed.

[0002]

2. Description of the Related Art For example, the following methods have been known as methods for producing cytidine derivatives. (1) 5'-O- (dimethoxytrityl) -uridine derivative is condensed with 1,2,4-triazole to give 1- [5 '
-O- (dimethoxytrityl) -β-D-ribofuranosyl] -4- (1,2,4-triazol-1-yl) pyrimidin-2- (1H) one derivative, which is then reacted with amines in dioxane solvent. Method for obtaining 5'-O- (dimethoxytrityl) cytidine derivative [Journal
of Organic Chemistry, 47,
3623 (1982)]. (2) 2′-deoxy-2′-methyluridine derivative is converted to 4- (N, N-dimethylamino) pyridine (hereinafter, DM
AP) to give the 4- [4- (N, N-dimethylamino) pyridinium] derivative, followed by 28%
Method for obtaining 2′-deoxy-2′-methylcytidine derivative with NH ₄ OH [Arch. Pharm. , 32
9, 66 (1996)].

In the above method (1) using 1,2,4-triazole, a 4- (1,2,4-triazol-1-yl) pyrimidin-2- (1H) one derivative is synthesized. Is very slow and takes a long time.
The product must be extracted, which is troublesome and unsuitable for mass production. The method of (2) using a 4- [4- (dimethylamino) pyridinium] derivative is described in a comparative example described below, but the reaction is extremely slow and requires a long time.
If the amount of AP used is less than 2.0 equivalents relative to the reaction substrate, the substrate will remain and a separation operation from the target substance will be required,
Because of the number of steps, it is not suitable for mass production.

[0004]

As described above, heretofore, there has been no method for producing a cytidine derivative that can be supplied efficiently and in large quantities. The present invention provides a production method for efficiently producing a cytidine derivative.

[0005]

Means for Solving the Problems The present inventors have made intensive studies to solve the above problems, and as a result, reacted a uridine derivative with a tertiary amine and a dehydrating reactant in the presence of a deoxidizing agent. Subsequently, it was found that when the cytidine derivative was reacted with ammonia or a primary or secondary amine, a cytidine derivative could be obtained in a short and easy operation. That is, the reaction time was long in the conventional method, but the reaction time was significantly shortened. Moreover
The tertiary amine can be reduced to almost equimolar to the uridine derivative, and the present invention has been completed. That is, the method for producing a cytidine derivative of the present invention is represented by formula (1)

[0006]

Embedded image

(Wherein X represents a hydrogen atom, a halogen atom, an alkyl group having 1 to 4 carbon atoms, an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, or an alkenyl group having 2 to 4 carbon atoms. , R2 each independently represent a hydrogen atom or a hydroxyl-protecting group; R3 represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, a cyano group, an alkenyl group, an alkynyl group,
Represents a hydroxyl group substituted with an alkoxy group and a hydroxyl-protecting group, and a tertiary amine and a dehydrating reactant.

[0008]

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(Wherein R 4 and R 5 are each independently a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 5 to 8 carbon atoms, or an alkyl having 1 to 4 carbon atoms substituted with a halogen atom) A alkenyl group having 2 to 4 carbon atoms, and R4 and R5 may be bonded to each other to form a ring), or a primary or secondary amine. , Equation (3)

[0010]

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(Wherein, X, R1, R2, R3, R4 and R5 have the same meanings as described above).

The hydroxyl-protecting groups as R1 and R2 in each of the above formulas include an aliphatic acyl group having 1 to 4 carbon atoms, an aromatic acyl group, and an aromatic group substituted with an alkyl group having 1 to 4 carbon atoms. An acyl group, an aromatic acyl group substituted with a halogen atom, an aromatic acyl group substituted with an alkoxy group having 1 to 4 carbon atoms, and a trialkylsilyl group represented by R3
A hydrogen atom, an alkoxy group having 1 to 4 carbon atoms, an aliphatic alkyloxy group having 1 to 4 carbon atoms substituted by an alkoxy group having 1 to 4 carbon atoms, an aliphatic acyloxy group having 1 to 4 carbon atoms, and aromatic An aromatic acyloxy group, an aromatic acyloxy group substituted with an alkyl group having 1 to 4 carbon atoms, an aromatic acyloxy group substituted with a halogen atom, and an aromatic acyloxy group substituted with an alkoxy group having 1 to 4 carbon atoms are preferable. Can be listed as

X and R3 in the above formulas (1) and (3)
A preferable combination is a combination wherein X represents a hydrogen atom or a methyl group, and R3 is a hydrogen atom, a methoxy group or a methoxyethyl group.

The tertiary amine is represented by the formula (4)

[0015]

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(Wherein, n and m each independently represent an integer of 1 to 4, and Y represents a hydrogen atom, a carbon atom, a nitrogen atom,
Z represents a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, and 1 carbon atom substituted by a halogen atom;
To 4 alkyl groups, alkenyl groups having 2 to 4 carbon atoms,
A represents a bond with A to form a ring, and A represents an alkyl group having 1 to 4 carbon atoms, an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, and alkenyl having 2 to 4 carbon atoms. Alicyclic amine represented by the formula (6):

[0017]

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(Wherein R6, R7 and R8 each independently represent an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 5 to 8 carbon atoms, or 1 carbon atom substituted by a halogen atom)
To 4 alkyl groups and alkenyl groups having 2 to 4 carbon atoms).

Specific examples of the tertiary amine include N-methylpiperidine, N-methylmorpholine, 1,4-diazabicyclo [2.2.2] octane, N, N'-dimethylpiperazine, and trimethylamine. Can be.

On the other hand, in the method for producing a cytidine derivative according to the present invention, when a uridine derivative is reacted with a tertiary amine and a dehydrating reactant, a compound represented by the formula (5)

[0021]

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(Where X, R1, R2, R3, n, m,
A, Y, and Z have the same meanings as described above).

As the dehydrating reactant in the above method, an acid halide or an acid anhydride can be mentioned. In this case, it is preferable to carry out the above reaction in the presence of a deoxidizing agent. Further, as a specific example of the above-mentioned dehydrating reactant, p-toluenesulfonic acid chloride can be mentioned.

The amount of the tertiary amine to be used relative to the uridine derivative represented by the formula (1) can be, for example, 1.2 times or less.

Equation (5)

[0026]

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(Where X, R1, R2, R3, n, m,
A, Y, and Z have the same meanings as described above), or a salt thereof is a novel compound and is included in the present invention. In the compound of the above formula (5), X represents a hydrogen atom or a methyl group, R1 and R2 are a hydrogen atom or a hydroxyl-protecting group, R3 is a hydrogen atom, a methoxy group, a methoxyethyloxy group, and n and m are 2 , A is a methyl group,
Compounds in which Y is a methylene group or an oxygen atom can be mentioned.

Another embodiment of the method for producing a cytidine derivative according to the present invention is characterized in that the cytidine derivative represented by the above formula (5) or a salt thereof is reacted with ammonia or a primary or secondary amine. Equation (3)

[0029]

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(Wherein, X, R1, R2, R3, R4 and R5 have the same meanings as described above).

[0031]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail.

X in the uridine derivative represented by the formula (1) (hereinafter referred to as uridine derivative (1)) represents a hydrogen atom, a halogen atom such as a fluorine atom, a chlorine atom, a bromine atom or an iodine atom; 1 to 4 alkyl groups, i.e., methyl, ethyl, propyl, 2-propyl, tertiary
Butyl group, an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, that is, a chloromethyl group, a dichloromethyl group,
A trifluoromethyl group, an alkenyl group having 2 to 4 carbon atoms, that is, a bromovinyl group, but is not limited thereto.

R1 and R2 of the uridine derivative (1) independently represent a hydrogen atom or a hydroxyl-protecting group. Examples of the hydroxyl-protecting group include alkyl ethers, aralkyls, acyls, carbonates, sulfonates, and silyls.

As the alkyl ethers, a methoxymethyl group, a 2-methoxyethyl group, a 1-ethoxyethyl group, a 2-methoxyethoxymethyl group, a tetrahydropyranyl group and the like can be used.

The aralkyls include a benzyl group,
A methoxybenzyl group, a 2-nitrobenzyl group, a trityl group, a 4-methoxytrityl group, a 4,4′-dimethoxytrityl group and the like can be used.

As the acyls, any of an aliphatic acyl group and an aromatic acyl group can be used. These acyl groups may have one or more substituents such as a halogen, an alkyloxy group, a nitro group, an acyl group, and an alkyl group, if necessary.

More preferably, the acyl group is an aliphatic acyl group such as an acetyl group or a propionyl group, a benzoyl group, a toluoyl group, a nitrobenzoyl group, a 4-chlorobenzoyl group, a 3-chlorobenzoyl group, or a 2-chlorobenzoyl group. And aromatic acyl groups such as a 4-methoxybenzoyl group.

As the carbonates, a methoxycarbonyl group, an ethoxycarbonyl group, a tertiary butyloxycarbonyl group, a benzyloxycarbonyl group, a phenyloxycarbonyl group and the like can be used.

As the sulfonates, a p-toluenesulfonyl group, a methanesulfonyl group, a 2,4,6-trimethylphenylsulfonyl group, a 2,4,6-triisopropylphenylsulfonyl group and the like can be used.

The silyls include a trimethylsilyl group,
Triethylsilyl group, dimethyl t-butylsilyl group, diphenylmethylsilyl group, triphenylsilyl group,
1,3,3-tetraisopropyldisiloxane-1,3
-A diyl group or the like can be used.

Examples of R3 of the uridine derivative (1) include a hydrogen atom, a halogen atom such as a fluorine atom, a chlorine atom, a bromine atom and an iodine atom, a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, ie, a methyl group, Ethyl group,
Alkenyl groups such as propyl group, 2-propyl group, tertiary butyl group, cyano group, and bromovinyl group; alkynyl groups such as ethynyl group; alkoxy groups having 1 to 4 carbon atoms, that is, methoxy group, ethyloxy group, and n-propyloxy group; , N-butyloxy group, or a hydroxyl group substituted with the above-mentioned hydroxyl protecting group. More preferably, they are a hydrogen atom, a methoxy group, and a methoxyethyloxy group.

The tertiary amines represented by the formulas (4) and (6) include, for example, trimethylamine, triethylamine, tripropylamine, tri (2-propyl) amine, tributylamine, tri (2-butyl) amine, tri ( number 3
Butyl) amine, di (2-propyl) ethylamine, N
-Methylpyrrolidine, N-methylpiperidine, N-methylpyrrole, N, N'-dimethylpiperazine, 1,4-
Examples thereof include diazabicyclo [2.2.2] octane, N-methylmorpholine, and N-methylthiomorpholine, but are not limited thereto as long as the nucleophilicity of the nitrogen atom is sufficient for the progress of the reaction.

Among them, trimethylamine, N-methylpyrrolidine, 1,4-diazabicyclo [2.2.2] octane, N-methylpiperidine, N-methylmorpholine and N, N'-dimethylpiperazine are particularly preferred.

Ammonia or 1 represented by the formula (2)
R4 and R5 of the secondary or secondary amine are independently a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, that is, a methyl group, an ethyl group, a propyl group, a 2-propyl group, a tertiary butyl group,
A cycloalkyl group having 5 to 8 carbon atoms, that is, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group, a carbon atom having 1 to 4 carbon atoms substituted with a halogen atom;
Alkenyl groups having 2 to 4 carbon atoms such as chloromethyl group, dichloromethyl group and bromovinyl group. Further, R4 and R5 may combine to form a ring, in which case R4 (R
5) The N group can be a pyrrolidine group or a piperidine group, but is not limited thereto. Among them, ammonia and piperidine are particularly preferred.

Examples of the dehydrating reactant include, but are not limited to, acid halides, acid anhydrides, ester / amidating agents, acid catalysts and fluorinating agents.

As the acid halide, p-toluenesulfonyl chloride, methanesulfonyl chloride, 2,2
4,6-trimethylphenylsulfonyl chloride, 2,
4,6-triisopropylphenylsulfonyl chloride, phosphorus oxychloride, thionyl chloride, dichlorophosphate 4-
Chlorophenyl, oxalyl chloride, malonyl dichloride and the like can be used. As the acid anhydride, trifluoromethanesulfonic anhydride, acetic anhydride, trifluoroacetic anhydride and the like can be used. Examples of the ester / amidating agent include carbodiimides such as 1,3-dicyclohexylcarbodiimide, phosphonium salts such as bromo (trispirolidino) phosphonium hexafluorophosphate, 2-chloro-N-methylpyridinium iodide and the like. And azocarboxylates such as diethyl azodicarboxylate. As the acid catalyst, boron trifluoride diethyl ether, tin tetrachloride, aluminum trichloride, or the like can be used. As the fluorinating agent, diethylaminosulfur trifluoride, cyanuric fluoride, 1,3-dimethyl-2,2-difluoroimidazoline and the like can be used. Acid halides are preferred, and p-toluenesulfonyl chloride is particularly preferred.

Examples of the reaction solvent for this reaction include ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran and dioxane, aliphatic hydrocarbon solvents such as n-pentane, n-hexane and cyclohexane, benzene, toluene, xylene and halogen. Aromatic hydrocarbon solvents such as halogenated benzene, halogenated hydrocarbon solvents such as dichloromethane, chloroform and dichloroethane, and acetonitrile can be used, but are not limited thereto. Of these, acetonitrile and chloroform are particularly preferred.

The reaction of the uridine derivative (1) of the present invention with a tertiary amine and a dehydrating agent is performed by adding a dehydrating agent or a solution thereof to a solution of the uridine derivative (1) and a tertiary amine. It is performed by adding.

When the acidity in the reaction system affects the reaction, a deoxidizing agent can be added separately from the tertiary amine. Examples of the deoxidizing agent include, in addition to the above-mentioned tertiary amine, organic bases such as pyridine, lutidine, N, N-dimethylaniline, inorganic bases such as potassium carbonate, sodium carbonate and sodium hydrogencarbonate, and ion exchange resins. And the like, but are not limited to these. Among them, triethylamine is particularly preferred.

The amount of the tertiary amine to be used relative to the uridine derivative (1) is in the range of 0.5 to 3.0 moles, preferably 1.0 to 2.0 moles.
Among them, a 1.2-fold molar amount is more preferred.

The amount of the dehydrating reactant used in the reaction of the uridine derivative (1) with the tertiary amine and the dehydrating reactant is 0.5 to 5.0 moles per mol of the uridine derivative (1). The molar ratio is from 1.0 to 3.0 times, preferably from 2.0 to 3.0 times, more preferably 2.0 times.

When the uridine derivative (1) is allowed to coexist with the tertiary amine and the dehydrating reactant during the reaction, the amount of the deoxidizing agent is 0.5 times the mol of the uridine derivative (1). To 5.0 moles, preferably 1.0 to 3.0 moles, and more preferably 2.0 moles.

The temperature at which the dehydrating reactants or their solutions are added to the solution of the uridine derivative (1) and the tertiary amine can be in the range of -10 ° C. to 50 ° C., but is preferably It is in the range of -5 ° C to 10 ° C.

The reaction time of the uridine derivative (1) with the tertiary amine and the dehydrating reactant is 0.5 to 2 hours.
It takes 7 hours, but preferably between 0.5 and 3 hours.

The synthesis of the cytidine derivative (3) of the present invention
The reaction is carried out by reacting the uridine derivative (1) with a tertiary amine and a dehydrating reactant, and then reacting with ammonia or primary and secondary amines. The amount of the ammonia or the primary and secondary amines (2) to be used in the uridine derivative (1) is in the range of 0.5 to 60 times, preferably in the range of 1.0 to 40 times, and more preferably in the range of 1.0 to 40 times. The range of 2.0 to 37 moles is more preferable.

After reacting the uridine derivative (1) with a tertiary amine and a dehydrating reactant, the reaction with ammonia or a primary or secondary amine is carried out at a temperature of -10 ° C to 50 ° C. C., but is preferably between -5.degree. C. and 10.degree. C. and takes between 0.5 and 10 hours, but preferably between 0.5 and 6 hours. In this reaction, the activation of the uridine derivative at the 4-position is first advanced by a dehydrating reagent, and the activated uridine derivative (1) is represented by the formula (5) by reaction with a tertiary amine. Cytidine derivative or formula (7)

[0057]

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(Wherein R1, R2, R3, R6, R7, R
8 and X have the same meanings as defined above), and then react with ammonia or primary and secondary amines to proceed to amination at the 4-position.

The cytidine derivative (3) can also be produced by isolating the cytidine derivatives (5) and (7) and treating them with ammonia or primary and secondary amines.

According to the present invention, the cytidine derivative (3) can be produced more efficiently.

[0061]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the invention is limited thereto.

Example 1 3 ', 5'-O-bis (4-chlorobenzoyl) -2'
-50 g of acetonitrile in 10.0 g of deoxyuridine
L, 4.0 g of triethylamine and 2.4 g of 1-methylpiperidine (1.2 times mol of the uridine derivative) were added,
Stir in an ice bath. A solution obtained by mixing 7.5 g of p-toluenesulfonyl chloride and 25 mL of acetonitrile was added dropwise at -1.5 ° C or lower over 1 hour. After completion of the dropwise addition, the mixture was stirred for 1 hour. Add 50 mL of 28% ammonia water to 4.5
The solution was added dropwise over 20 minutes at a temperature of not more than ° C. 2 hours after dropping 30
Minutes. The precipitated crystals were filtered and washed with acetonitrile to obtain 7.93 g of 3 ', 5'-O-bis (4-chlorobenzoyl) -2'-deoxycytidine. Yield 79.5%.

¹ H-NMR (400 MHz, DMSO-d
6) δ 2.47 to 2.55 (m, 2H), 3.35
(S, 1H), 4.50-4.63 (m, 3H), 5.
59-5.61 (m, 1H), 5.70 (d, J = 7.
6 Hz, 1 H), 6.29 (t, J = 7.0 Hz, 1
H), 7.23 (d, J = 7.8 Hz, 1H), 7.5
9 to 7.66 (m, 5H), 7.96 to 8.04 (m, 5H)
4H). IR (KBr) cm ^-1 1719, 1655, 149
1,1271,1095. Comparative Example 1 The method shown in the conventional method (2) using DMAP was additionally tested. It is described below. 0.283 g of 3 ', 5'-O-bis (4-chlorobenzoyl) -2'-deoxyuridine
To this were added 8.5 mL of acetonitrile, 0.221 g of p-toluenesulfonyl chloride and 0.141 g of DMAP (2.1 times the mol of the uridine derivative), and 0.117 g of triethylamine was added dropwise, followed by stirring at room temperature for 22 hours. 5.7 mL of 28% ammonia water was added thereto, and the mixture was stirred at room temperature for 2 hours and cooled with ice. The crystals were filtered and washed with 60% acetonitrile-water to obtain 0.17 g of 3 ', 5'-O-bis (4-chlorobenzoyl) -2'-deoxycytidine. Yield 60%.

As described above, in the conventional method, it is necessary to stir at room temperature for 22 hours before adding 28% aqueous ammonia, but in the method of the present invention, it is sufficient to stir for 1 hour, and the reaction time can be greatly reduced. Was.

Comparative Example 2 An experiment was conducted in which the number of equivalents of DMAP was changed. It is described below.

3.0 ml of acetonitrile and 0.2 ml of p-toluenesulfonyl chloride were added to 0.2 g of 3 ', 5'-O-bis (4-chlorobenzoyl) -2'-deoxyuridine.
151 g and the equivalent number of DMAP shown in Table 1 were added, and 80.2 mg of triethylamine was added dropwise, and 30
Stirred at room temperature for hours. The reaction solution was analyzed by high performance liquid chromatography, and the reaction substrate 3 ′, 5′-O-bis (4
-Chlorobenzoyl) -2'-deoxyuridine was determined. Table 1 shows the results.

[0067]

[Table 1]

As described above, when DMAP is 1.2 equivalents or less, the raw material remains, and a separation operation from the target substance is required, and the number of steps is increased. This is probably because DMAP is deactivated by hydrogen chloride generated in the reaction system.
On the other hand, the tertiary amine shown in the present invention is not deactivated by hydrogen chloride, and the target substance can be obtained in a short time without leaving any reaction substrate even when 1.2 equivalents are used.

Example 2 3 ', 5'-O-bis (4-chlorobenzoyl) -2'
-5 g of acetonitrile in 1.0 g of deoxyuridine,
0.40 g of triethylamine, 0.24 g of 1-methylpiperidine (1.2 times mol of uridine derivative) and p-
0.76 g of toluenesulfonyl chloride was added, and the mixture was stirred in an ice bath for 2 hours. 7.5 mL of 2-propanolic ammonia was added thereto, and the mixture was stirred in an ice bath for 4 hours.
Stirred for hours. The crystals are filtered, washed with acetonitrile and 3 ', 5'-O-bis (4-chlorobenzoyl)-
0.54 g of 2'-deoxycytidine was obtained. Yield 54
%.

Example 3 3 ', 5'-O-bis (4-chlorobenzoyl) -2'
-2.0 g of deoxyuridine in 60 m of acetonitrile
L, 0.81 g of triethylamine, 0.54 g of 1,4-diazabicyclo [2.2.2] octane (1.2 times mol of uridine derivative) and 1.5 g of p-toluenesulfonyl chloride were added, and the mixture was stirred at room temperature for 3 hours. did. Ammonia gas was blown into the mixture at room temperature for 1 hour, followed by stirring in an ice bath for 30 minutes. Filter the crystals, wash with acetonitrile,
3 ', 5'-O-bis (4-chlorobenzoyl) -2'
-1.52 g of deoxycytidine were obtained. Yield 75.2
%.

Example 4 3 ', 5'-O-bis (4-chlorobenzoyl) -2'
-30 g of chloroform in 3.0 g of deoxyuridine,
0.72 g of triethylamine, 0.80 g of 1,4-diazabicyclo [2.2.2] octane (1.2 times mol of uridine derivative) and 1.36 g of p-toluenesulfonyl chloride were added, and the mixture was stirred at room temperature for 30 minutes. Ammonia gas was blown into the mixture at room temperature for 1 hour, followed by stirring in an ice bath for 1 hour. After adding 100 mL of water and 100 mL of methanol to sludge, the crystals are filtered, washed with a mixed solvent of water and methanol (1: 1), and 3 ′, 5′-O-bis (4-chlorobenzoyl) -2. 2.42 g of '-deoxycytidine were obtained. Yield 80.9%.

Reference Example 1 5.0 g of thymine was suspended in 41.8 mL of HMDS.
Heated to reflux for an hour. After cooling, excess HMDS was distilled off. After adding 60 mL of chloroform to the residue and dissolving,
3,5-O-bis (4-chlorobenzoyl) -2-deoxyribofuranose-1-yl chloride 11.4 g
(85% content), 60 mL of chloroform was further added, and the mixture was heated and stirred at 50 ° C. for 4 hours. After cooling, a solution of 4.46 g of sodium hydrogencarbonate in 70 mL of water and 70 mL of methanol were added, followed by stirring at room temperature for 1 hour. After removing the aqueous layer, a solution of 300 mg of sodium carbonate in 30 mL of water was added, and the mixture was stirred at room temperature for 10 minutes. After liquid separation, the solvent of the organic layer was distilled off, and the obtained residue was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to give 3 ′, 5′-O-bis (4-chlorobenzoyl). 1) 10.5 g of thymidine were obtained. 90% yield.

¹ H-NMR (400 MHz, CDCl ₃ )
δ 1.65 (d, J = 0.7 Hz, 3H), 2.55
-2.66 (m, 2H), 4.48-4.68 (m, 3
H), 5.61-5.65 (m, 1H), 6.28-
6.32 (m, 1H), 7.50 (d, J = 0.7H)
z, 1H), 7.57-7.64 (m, 4H), 7.9.
9-8.04 (m, 4H), 11.4 (s, 1H). IR (KBr) cm ^-1 3193, 3067, 17
20, 1680, 1593, 1488, 127
6, 1013, 761. Example 5 3 ', 5'-O-bis (4-chlorobenzoyl) thymidine (uridine derivative in which X is a methyl group) In a suspension of 3.00 g of acetonitrile in 15 mL of 1-methylpiperidine 0.85 mL (uridine derivative (1.2 times the mol of) and 1.70 mL of triethylamine, and then cooled.
A solution of 2.31 g of p-toluenesulfonyl chloride in 15 mL of acetonitrile was added dropwise while maintaining the temperature at 0 ° C or lower, and the mixture was stirred for 3 hours while maintaining the temperature at 0 ° C or lower. While maintaining the temperature at 0 ° C. or lower, 15 mL of 28% aqueous ammonia was added thereto, followed by stirring for 6 hours. The solvent was filtered and the residue
0 mL was added, and the mixture was stirred under ice cooling for 2 hours. The solvent was filtered, and the obtained solid was dried to obtain 1.59 g of 3 ′, 5′-O-bis (4-chlorobenzoyl) -5-methyl-2′-deoxycytidine as a light brown solid. Yield 52%.

¹ H-NMR (400 MHz, DMSO-
d6) δ 1.68 (s, 3H), 2.42-2.56
(M, 2H), 4.47-4.50 (m, 1H), 4.
54-4.59 (m, 1H), 4.64-4.69
(M, 1H), 5.60-5.64 (m, 1H), 6.
31-6.35 (m, 1H), 6.86 (brs, 1
H), 7.39 (brs, 2H), 7.59-7.6.
4 (m, 4H), 7.98-8.04 (m, 4H). IR (KBr) cm ^-1 3470, 1681, 148
8, 1270, 1093, 760. Reference Example 2 3 ', 5'-O-bis (4-chlorobenzoyl) -5
To a suspension of 0.71 g of methyl-2'-deoxycytidine in 4 mL of methanol was added 0.7 mL of a sodium hydroxide-methanol solution (adjusted by dissolving 100 mg of sodium hydroxide in 4 mL of methanol), and heated at 45 ° C for 5 hours. Stirred. After cooling, the mixture was neutralized with a hydrochloric acid-methanol solution, and methanol was distilled off. After adding chloroform and water to the residue, liquid separation was performed, and the aqueous layer was washed with chloroform. After concentrating the aqueous layer, a 6N aqueous hydrochloric acid solution was added to make the mixture acidic, 5 mL of acetone was added, and the mixture was allowed to stand at -14 ° C overnight. The precipitated solid was collected by filtration and dried to obtain 0.37 g of 5-methyl-2′-deoxycytidine hydrochloride as a light brown solid. 97% yield.

Reference Example 3 To a solution of 1.01 g of 2'-O-methyluridine and 0.67 g of imidazole in 20 mL of DMF was added 1.3 g of chloro (t-butyl) dimethylsilane, and the mixture was stirred at room temperature for 7 hours. After completion of the reaction, the mixture was concentrated, and the residue was subjected to silica gel column chromatography (ethyl acetate: n-hexane = 3:
Purified in 7) and 1.44 g of 3 ', 5'-O-bis (t-butyldimethylsilyl) -2'-O-methyluridine
Was obtained as a colorless powder. Yield 76%.

¹ H-NMR (400 MHz, DMSO-
d6) δ 0.085 (s, 3H), 0.10 (s, 3
H), 0.12 (s, 3H), 0.13 (s, 3H),
0.91 (s, 9H), 0.94 (s, 9H), 3.5
5 (s, 3H), 3.60 (dd, J = 1.7 & 4.9)
Hz, 1H), 3.77 (dd, J = 2.0 & 12.0)
Hz, 1H), 4.18 (dd, J = 2.0 & 12.0)
Hz, 1H), 4.02 to 4.06 (m, 1H), 4.
24 (dd, J = 4.9 & 7.1 Hz, 1H), 5.6
8 (dd, J = 2.0 & 8.3 Hz, 1H), 5.94
(D, J = 2.0 Hz, 1H), 8.07 (d, J =
8.3 Hz, 1H), 8.77 (brs, 1H). IR (KBr) cm ^-1 3463,2953,293
0,2858,1691,1630,1542,146
3,1192,1124,1035,1012,83
9,683. Example 6 3 ', 5'-O-bis (t-butyldimethylsilyl)-
0.23 mL of 1-methylpiperidine was added to a solution of 750 mg of 2′-O-methyluridine in 10 mL of acetonitrile.
(1.2 times mol of uridine derivative) and 0.45 mL of triethylamine were added, and the mixture was cooled. A solution of 614 mg of p-toluenesulfonyl chloride in 5 mL of acetonitrile was added dropwise under ice-cooling, and the mixture was stirred for 1 hour. After the disappearance of the starting materials, 1 mL of the reaction solution was taken and concentrated, and diethyl ether was added to the residue, and the precipitated crystals were removed by filtration. After concentrating the filtrate,
The residue was washed with n-hexane, dried, and [[3,5-O-
Bis (t-butyldimethylsilyl) -2-O-methylribofuranose-1-yl] -2-oxo-1,2-dihydro-4-pyrimidinyl] -1-methylpiperidinium chloride (reaction intermediate)

[0077]

Embedded image

Was obtained as a pale yellow oil.

¹ H-NMR (400 MHz, DMSO-
d6) δ 0.065 (s, 3H), 0.088 (s,
3H), 0.095 (s, 3H), 0.10 (s, 3
H), 0.89 (s, 18H), 2.2 to 1.6 (m,
6H), 3.65 (s, 3H), 3.69 (s, 3
H), 3.74 (d, J = 4.4 Hz, 1H), 3.8
0 (d, J = 12.0 Hz, 1H), 4.0 to 4.2
(M, 5H), 4.60 (m, 2H), 5.92 (s,
1H), 7.40 (d, J = 7.3 Hz, 1H), 8.
88 (d, J = 7.3 Hz, 1H). IR (KBr) cm ^-1 3453, 3194, 305
8,2953,2930,2859,1702,146
2,1256,1129,1073,838,780. The remaining reaction solution was cooled under ice-cooling, 28% ammonia water 3.5 m
L was added and stirred for 2 hours. The reaction solution was concentrated, diluted with ethyl acetate, and washed with water and saturated saline. The organic layer was dried over magnesium sulfate, concentrated, and the residue was purified by silica gel column chromatography (methanol: chloroform = 1: 25) to give 3 ', 5'-O-bis (t-
Butyldimethylsilyl) -2′-O-methylcytidine 5
44 mg were obtained as a colorless powder. Yield 73%.

¹ H-NMR (400 MHz, DMSO-
d6) δ 0.054 (s, 3H), 0.076 (s,
3H), 0.11 (s, 3H), 0.13 (s, 3
H), 0.89 (s, 9H), 0.94 (s, 9H),
3.61 (d, J = 4.9 Hz, 1H), 3.64
(S, 3H), 3.78 (d, J = 12.0 Hz, 1
H), 4.04 (d, J = 9.0 Hz, 1H), 4.0.
9 (d, J = 12.0 Hz, 1H), 4.18 (dd,
J = 4.9 & 8.8 Hz, 1H), 5.61 (d, J =
7.3 Hz, 1H), 5.93 (s, 1H), 8.17
(D, J = 7.3 Hz, 1H). IR (KBr) cm ^-1 3355, 3200, 295
5,2930,2859,1647,1491,125
5,1129,1073,838,781.

[0081]

According to the present invention, a cytidine derivative can be produced more efficiently by using a method which can be mass-produced than by a conventional method.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Toshiyuki Kono 1144 Togo, Togo, Mobara-shi, Chiba Mitsui Chemicals, Inc. (72) Inventor Hironori Komatsu 1144, Togo, Togo, Mobara-shi, Chiba Mitsui Chemicals, Inc. (72) Inventor, Nobuyuki Fukasawa 1144 Togo, Mobara-shi, Chiba Mitsui Chemicals, Inc. F-term (reference) 4C057 BB02 DD01 LL10 LL19

Claims (13)

[Claims] (1) Formula (1) (Wherein X is a hydrogen atom, a halogen atom, an alkyl group having 1 to 4 carbon atoms, and a carbon atom having 1 to 4 carbon atoms substituted with a halogen atom)
Represents an alkenyl group having 2 to 4 carbon atoms,
R1 and R2 each independently represent a hydrogen atom or a hydroxyl-protecting group; R3 represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group having 1 to 4 carbon atoms, a cyano group, an alkenyl group, an alkynyl group, and a carbon atom; (Representing a hydroxyl group substituted with an alkoxy group of 1 to 4 and a hydroxyl-protecting group)) with a tertiary amine and a dehydrating reactant, and then reacting with a compound of the formula (2) (Wherein R4 and R5 each independently represent a hydrogen atom, an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 5 to 8 carbon atoms, an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, Alkenyl groups of the formulas 2 to 4, R4 and R
5 represents that it may be bonded to form a ring), or a primary or secondary amine represented by the formula (3): (Wherein, X, R1, R2, R3, R4 and R5 have the same meanings as described above). 2. R1 and R2 each independently have 1 carbon atom
To 4, an aliphatic acyl group, an aromatic acyl group, an aromatic acyl group substituted with an alkyl group having 1 to 4 carbon atoms, an aromatic acyl group substituted with a halogen atom, and an alkoxy group having 1 to 4 carbon atoms. A substituted aromatic acyl group or a trialkylsilyl group, wherein R 3 is a hydrogen atom, an alkoxy group having 1 to 4 carbon atoms, or an aliphatic alkyl having 1 to 4 carbon atoms substituted with an alkoxy group having 1 to 4 carbon atoms; Oxy group, aliphatic acyloxy group having 1 to 4 carbon atoms, aromatic acyloxy group, aromatic acyloxy group substituted with an alkyl group having 1 to 4 carbon atoms, aromatic acyloxy group substituted with a halogen atom, 1 carbon atom The method for producing a cytidine derivative according to claim 1, wherein the aromatic oxy group is an aromatic acyloxy group substituted with an alkoxy group. 3. X represents a hydrogen atom or a methyl group;
The method for producing a cytidine derivative according to claim 2, wherein R3 is a hydrogen atom, a methoxy group, or a methoxyethyl group. 4. A tertiary amine of the formula (4) (In the formula, n and m each independently represent an integer of 1 to 4, Y represents a hydrogen atom, a carbon atom, a nitrogen atom, an oxygen atom, or a sulfur atom, and Z represents a hydrogen atom, an alkyl having 1 to 4 carbons. Group, an alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, an alkenyl group having 2 to 4 carbon atoms, which may be bonded to A to form a ring, and A represents 1 to 4 carbon atoms. An alkyl group having 1 to 4 carbon atoms substituted with a halogen atom, an alkenyl group having 2 to 4 carbon atoms, Z
The method for producing a cytidine derivative according to any one of claims 1 to 3, wherein the alicyclic amine is represented by the following formula: 5. A reaction intermediate obtained by reacting a uridine derivative with a tertiary amine and a dehydrating reactant is represented by the formula (5): (Wherein X, R1, R2, R3, n, m, A, Y, and Z have the same meanings as described above), wherein the cytidine derivative according to claim 4, wherein Manufacturing method. 6. A tertiary amine of the formula (6) (Wherein, R6, R7 and R8 each independently represent an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 5 to 8 carbon atoms, an alkyl group having 1 to 4 carbon atoms substituted by a halogen atom, The method for producing a cytidine derivative according to any one of claims 1 to 3, wherein the amine is an aliphatic amine represented by formula (2). 7. The tertiary amine is N-methylpiperidine, N
-Methylmorpholine, 1,4-diazabicyclo [2.
2.2] The method for producing a cytidine derivative according to any one of claims 1 to 6, which is octane, N, N'-dimethylpiperazine, or trimethylamine. 8. The method for producing a cytidine derivative according to claim 1, wherein the dehydrating reactant is an acid halide or an acid anhydride, and the reaction is carried out in the presence of a deoxidizing agent. 9. The method for producing a cytidine derivative according to claim 1, wherein the dehydrating reactant is p-toluenesulfonic acid chloride. 10. The method for producing a cytidine derivative according to claim 1, wherein the amount of the tertiary amine to be used is 1.2 times or less the amount of the uridine derivative represented by the formula (1). 11. The formula (5) (Wherein, X, R1, R2, R3, n, m, A, Y, and Z are as defined above) or a salt thereof. 12. X represents a hydrogen atom or a methyl group, and R1 and R2 represent a hydrogen atom or a hydroxyl-protecting group;
3 is a hydrogen atom, a methoxy group, a methoxyethyloxy group,
The cytidine derivative or a salt thereof according to claim 11, wherein n and m are 2, A is a methyl group, and Y is a methylene group or an oxygen atom. 13. A cytidine derivative or a salt thereof according to claim 11 or 12, comprising ammonia or a primary or secondary compound.
A compound of the formula (3), which is reacted with a secondary amine. (Wherein, X, R1, R2, R3, R4 and R5 have the same meanings as described above).